Methods and devices for localized disease treatment by ablation

ABSTRACT

Provided herein are methods, systems, and devices for increasing heat shock protein expression and treating conditions for which increased heat shock protein expression is expected to be beneficial using thermal ablation.

RELATED APPLICATIONS

This application is a Divisional of U.S. patent application Ser. No.15/816,954, filed Nov. 17, 2017, now U.S. Pat. No. 11,020,176, which isa Continuation of and claims priority to U.S. patent application Ser.No. 14/390,962, filed Oct. 6, 2014, now U.S. Pat. No. 9,848,950, whichis a U.S. National Phase under 35 U.S.C. 371 of InternationalApplication No. PCT/US2013/030016, filed Mar. 8, 2013, which claimspriority to U.S. Provisional Patent Application No. 61/639,798, filedApr. 27, 2012, the disclosures of which are incorporated by referenceherein in their entirety.

BACKGROUND

Heat shock proteins (HSPs) (also known as stress proteins or molecularchaperones) are an evolutionarily conserved and diverse group ofmolecular chaperones that are upregulated in response to cellularstressors such as oxidative stress, glucose deprivation, hyperthermia,hypothermia, infection, inflammation, dehydration, ischemia, andexposure to toxins.

HSPs may be broadly classified into five families according to theirmolecular mass: HSP100, HSP90, HSP70, HSP60, and small HSPs (sHSPs).sHSPs are known to be abundant in cardiac and skeletal muscle, wherethey increase in response to stress to protect against muscle ischemia.

The chaperone activities of HSPs include prevention of proteinmisfolding, refolding of denatured proteins, and targeting of proteinsfor proteolytic degradation. In humans, HSPs are most notably associatedwith the cardiovascular, renal, central nervous, lymphatic, and immunesystems. In addition, there is a strong functional relationship betweenthe sympathetic nervous system (SNS) and HSPs.

HSPs play a key role in cell survival through cytoprotective mechanisms(Almeida Biomed Pharmacother 65:239 (2011)), and are involved inresponding to a variety of disease processes including those of cancer,cardiovascular disease, neurodegenerative disease, trauma, diabetes, andchronic inflammation.

BRIEF DESCRIPTION OF DRAWINGS

FIGS. 1A-1C: Examples of HSP detection methods: antibody-based detection(FIGS. 1A and 1B) and activity-based detection (FIG. 1C).

FIGS. 2A-2B: Illustrative digital (FIG. 2A) and analog (FIG. 2B) outputsfor displaying a detectable signal generated by the interaction of HSPswith a capture or detection agent.

FIG. 3 : Quantum dot system for generation of a detectable signalfollowing binding of an HSP to an affinity ligand capture agent.

FIGS. 4A-4B: Illustrative HSP capture methods: removal from ablationsite and sequestration in a capture compartment for analysis in vivo orex vivo (FIG. 4A), balloon-based/semi-permeable filtering device withantibody based/immuno-electrochemical technology embedded within forcapture and analysis in vivo or ex vivo (FIG. 4B).

FIG. 5 : Average kidney NE levels post-ablation.

FIG. 6 : Upregulation of HSPA5 24 hours post-ablation.

FIG. 7 : Upregulation of DNAJA4 10 minutes post-ablation.

FIG. 8 : Upregulation of DNAJB1 10 minutes post-ablation.

FIGS. 9A-9B: Upregulation of CLU (FIG. 9A) 10 minutes and (FIG. 9B) 7days post-ablation.

FIG. 10 : Upregulation of HSPD1 10 minutes, 24 hours, and 7 dayspost-ablation.

FIGS. 11A-11B: Upregulation of HSPH1 (FIG. 11A) 10 minutes and (FIG.11B) 24 hours post-ablation.

FIGS. 12A-12B: Upregulation of HMOX1 (FIG. 12A) 24 hours and (FIG. 12B)7 days post-ablation.

FIGS. 13A-13B: Illustration of representative electrochemicalimmunosensor protocols.

FIG. 14 : Embodiments of a system for carrying out the methods disclosedherein.

DETAILED DESCRIPTION

The present technology is directed to methods, systems, devices, andkits for increasing HSP expression levels at or near a target site usingablation, as well as to methods, systems, devices, and kits forincreasing local expression of HSP and treating conditions for whichincreased HSP levels are expected to be beneficial by increasing HSPexpression. Although many of the embodiments are described with respectto methods, systems, devices, and kits for treating various conditionsusing thermal ablation, other applications (e.g., use of non-thermalablative modalities) and other embodiments in addition to thosedescribed herein are within the scope of the technology. Additionally,several other embodiments of the technology can have differentconfigurations, components, or procedures than those described herein. Aperson of ordinary skill in the art, therefore, will accordinglyunderstand that the technology can have other embodiments withadditional elements, or the technology can have other embodimentswithout several of the features shown and described below.

Decreased induction of HSPs has been implicated in a variety ofdiseases, including cardiovascular disease (CVD) (e.g., myocardialinfarction (MI) or myocardial ischemia), neurodegeneration, trauma,stroke, diabetes, atherosclerosis, chronic inflammation, cancer, andvarious neurological disorders. Given the key role of HSPs incytoprotective mechanisms and the association between decreased HSPexpression and various disease states, HSPs represent attractivetherapeutic targets. Several attempts have been made to utilize HSPs astherapeutic agents. For example, whole-body hyperthermia (to induce thegeneral production of endogenous HSPs) and vaccination (to introduceamounts of specific exogenous HSPs) have been used to treat conditionsfor which increased HSP expression is expected to be beneficial. Despitepositive study results, targeted treatment has proven difficult. Thus,there is a need for new HSP treatment modalities for increasing HSPconcentration in a localized manner, for example to target treatment toa specific organ or system.

Thermal ablation is frequently used to remove or inactivate a targetbody tissue by applying targeted heat (hyperthermal ablation) or cold(hypothermal ablation). Examples of hyperthermal ablation techniquesinclude the use of monopolar or bipolar radio frequency (RF) energy,microwave energy, laser light or optical energy, ultrasound energy(e.g., intravascularly delivered ultrasound, extracorporeal ultrasound,high frequency ultrasound (HIFU)), magnetic energy, and direct heatenergy. Examples of hypothermal ablation techniques include the use ofcryotherapeutic energy. Hyperthermal ablation has been used to treat avariety of conditions, including cancer of the lung, liver, kidney, andbone, and conditions associated with electrical conduction in the heartsuch as arrhythmia.

Thermal ablation can also be used to partially or completely disrupt theability of a nerve to transmit a signal. For example, intravasculardevices that reduce sympathetic nerve activity by applying RF energy toa target site in the renal artery have recently been shown to reduceblood pressure in patients with treatment-resistant hypertension. Therenal sympathetic nerves arise from T10-L2 and follow the renal arteryto the kidney. The sympathetic nerves innervating the kidneys terminatein the blood vessels, the juxtaglomerular apparatus, and the renaltubules. Stimulation of renal efferent nerves results in increased reninrelease (and subsequent renin-angiotensin-aldosterone system (RAAS)activation) and sodium retention and decreased renal blood flow. Theseneural regulation components of renal function are considerablystimulated in disease states characterized by heightened sympathetictone and likely contribute to increased blood pressure in hypertensivepatients. The reduction of renal blood flow and glomerular filtrationrate as a result of renal sympathetic efferent stimulation is likely acornerstone of the loss of renal function in cardio-renal syndrome(i.e., renal dysfunction as a progressive complication of chronic heartfailure).

As disclosed herein, thermal ablation of renal sympathetic nerves hasbeen found to unexpectedly increase local expression of several HSPs.This increase was observed at 10 minutes, 24 hours, and 7 dayspost-ablation, indicating that HSP levels remain elevated for anextended period following ablation. Based on this finding, methods,systems, and devices are provided herein for increasing HSP expressionand for treating various conditions for which increased HSP levels areexpected to be beneficial.

Specific details of several embodiments of the technology are describedbelow with reference to FIGS. 1-14 . Although many of the embodimentsare described below with respect to methods, systems, and devices forinducing HSP expression and treating conditions for which increased HSPexpression is expected to be beneficial using thermal ablation, otherapplications (e.g., using other techniques such as non-thermalneuromodulation to increase HSP expression) and other embodiments inaddition to those described herein are within the scope of thetechnology. Additionally, several other embodiments of the technologycan have different configurations, components, or procedures than thosedescribed herein. A person of ordinary skill in the art, therefore, willaccordingly understand that the technology can have other embodimentswith additional elements, or the technology can have other embodimentswithout several of the features shown and described below with referenceto FIGS. 1-14 .

Disclosed herein are several embodiments of methods for increasing HSPexpression at or near a target tissue site and treating conditions forwhich increased HSP levels are expected to be beneficial in a subjectneed thereof using thermal ablation. The methods disclosed hereinrepresent a significant improvement over conventional approaches andtechniques in that they allow for more specific targeting of HSPinduction and thus localized treatment. Further provided herein aresystems and devices for use in conjunction with the disclosed methods.

Provided herein in certain embodiments are methods for increasing HSPexpression levels at or near a target site in a subject by performing athermal ablation procedure at or near the target site. In certain ofthese embodiments, HSP expression levels are increased within 30 minutesfollowing the thermal ablation procedure, and in certain of theseembodiments HSP expression levels are increased within 10 minutesfollowing the thermal ablation procedure. In certain embodiments, HSPexpression is increased at one or more specified timepoints followingablation, and in certain of these embodiments HSP expression isincreased at 10 minutes, 24 hours, or 7 days following ablation. Incertain embodiments, the thermal ablation procedure is carried out usingmonopolar or bipolar RF energy, microwave energy, laser light or opticalenergy, ultrasound energy (e.g., intravascularly delivered ultrasound,extracorporeal ultrasound, HIFU), magnetic energy, direct heat energy,or cryotherapeutic energy. In certain embodiments, the target site is ina renal blood vessel, and in certain of these embodiments the thermalablation procedure targets renal sympathetic nerves.

Provided herein in certain embodiments are methods for treatingconditions for which increased HSP expression is expected to bebeneficial in a subject by performing a thermal ablation procedure,wherein the procedure increases HSP expression levels. In certain ofthese embodiments, HSP expression levels are increased within 30 minutesfollowing the thermal ablation procedure, and in certain of theseembodiments HSP expression levels are increased within 10 minutesfollowing the thermal ablation procedure. In certain embodiments, HSPexpression is increased at one or more specified timepoints followingablation, and in certain of these embodiments HSP expression isincreased at 10 minutes, 24 hours, or 7 days following ablation. Incertain embodiments, the thermal ablation procedure is carried out usingmonopolar or bipolar RF energy, microwave energy, laser light or opticalenergy, ultrasound energy (e.g., intravascularly delivered ultrasound,extracorporeal ultrasound, HIFU), magnetic energy, direct heat energy,or cryotherapeutic energy. In certain embodiments, the target site is ina renal blood vessel, and in certain of these embodiments the thermalablation procedure targets renal sympathetic nerves.

Provided herein in certain embodiments are devices, systems, and kitsfor carrying out the methods disclosed herein.

In certain embodiments, the methods provided herein comprise performingthermal ablation, thereby increasing HSP expression at or near thetarget site. An increase in HSP expression may refer to an increase inmRNA or protein level for one or more HSPs or to an increase insecretion of one or more HSPs. In certain embodiments, thermal ablationresults in an increase in HSP activity at or near a target site inaddition to or in lieu of an increase in HSP expression. In certainembodiments, thermal ablation may be repeated one or more times atvarious intervals until a desired HSP expression level or anothertherapeutic benchmark is reached. An increase in HSP expression may beobserved for a single HSP, or it may observed across a range of two ormore HSPs. In certain embodiments, the methods disclosed herein resultin an increase in a single specific HSP. In other embodiments, themethods disclosed herein result in an increase in expression of a subsetof two or more specific HSPs, or an increase in HSP expressiongenerally. Examples of HSPs that may exhibit increased expression inresponse to thermal ablation are set forth in Table 1.

TABLE 1 Gene Product Gene Location Function/Description ReferenceClusterin (CLU) Secreted Secreted chaperone (heatshock Lu Curr Med Chem17:957 protein) (2010) DnaJ (Hsp40) homolog Intracellular/ Proteinfolding and heat response Sonna J Appl Physiol subfamily A member 4Surface 92:1725 (2002) (DNAJA4) DnaJ (Hsp40) homolog IntracellularInteracts with Hsp70, stimulates Lu Curr Med Chem 17:957 subfamily Bmember 1 ATPase activity (2010) (DNAJB1) Heat shock 27 kDa protein 1Intracellular Stress resistance, actin Lu Curr Med Chem 17:957 (HSPB1)organization (2010) Heat shock 60 kDa protein 1 IntracellularChaperonin, involved in folding of Lu Curr Med Chem 17:957 (HSPD1)mitochondrial matrix proteins (2010) Heat shock 105 kDa/110 kDaIntracellular Prevents aggregation of denatured Lu Curr Med Chem 17:957protein 1 (HSPH1) proteins under severe stress (2010) Heme oxygenase(decycling) 1 Intracellular Catalyzes degradation of heme, Sonna J ApplPhysiol (HMOX1) active during physiological stress 92:1725 (2002) Heatshock 70 kDa protein 5 Intracellular Facilitates assembly of multimericSABiosciences RT₂ Profiler (HSPA5) protein complexes in ER PCR ArrayHuman Neurotoxicity platform

In certain embodiments of the methods provided herein, thermal ablationresults in an increase in HSP levels over a specific timeframe. Incertain of these embodiments, HSP levels are increased over an acutetimeframe, e.g., within 30 minutes or less following ablation. Forexample, HSP levels may be increased at 30 seconds, 1 minute, 2 minutes,5 minutes, 10 minutes, 15 minutes, 20 minutes, or 30 minutespost-ablation. In certain embodiments, HSP levels may be elevated at oneor more timepoints beyond an acute timeframe. In these embodiments, HSPlevels may be elevated at or near the target site at 1 hour or more, 2hours or more, 6 hours or more, 12 hours or more, 24 hours or more, 48hours or more, 72 hours or more, or a week or more after the ablationprocedure. In certain embodiments where HSP levels are increased over anacute timeframe, levels remain increased beyond the acute timeframe. Inother embodiments, HSP levels are elevated during the acute timeframe,but drop back to a baseline level or below before the end of the acutetimeframe.

In certain embodiments, the methods disclosed herein may comprise anadditional step of measuring HSP expression levels, and in certain ofthese embodiments the methods further comprise comparing the HSPexpression level to a baseline HSP expression level. Such measurementsand comparisons can be used to monitor therapeutic efficacy and todetermine when and if to repeat the ablation procedure. In certainembodiments, a baseline HSP expression level is derived from the subjectundergoing treatment. For example, baseline HSP expression may bemeasured in the subject at one or more timepoints prior to thermalablation. Baseline HSP value may represent HSP expression at a specifictimepoint before thermal ablation, or it may represent an averageexpression level at two or more timepoints prior to thermal ablation. Incertain embodiments, the baseline value is based on HSP expression at ornear the target site immediately prior to thermal ablation (e.g., afterthe subject has already been catheterized). Alternatively, a baselinevalue may be derived from a standard value for HSP expression observedacross the population as a whole or across a particular subpopulation.In certain embodiments, post-ablation HSP levels are measured in anacute timeframe, i.e., while the subject is still catheterized and/orunder anesthesia.

In certain embodiments of the methods provided herein, the increase inHSP expression level post-ablation may result in an expression level ator near the target site that is greater (e.g., two-fold or greater,three-fold or greater, or five-fold or greater, etc.) than a baselineHSP expression level at one or more timepoints post-ablation. Forexample, HSP levels may be elevated to a level two-fold or greater thanthe baseline expression level at one or more timepoints within an acutetimeframe, e.g., within 30 minutes or less following ablation.

In certain embodiments of the methods provided herein, the methods aredesigned to increase HSP expression to a target level. In theseembodiments, the methods include a step of measuring HSP levelspost-ablation and comparing the resultant expression level to a baselineexpression level as discussed above. In certain of these embodiments,the ablation step is repeated until the target HSP level is reached. Inother embodiments, the methods are simply designed to increase HSP abovea baseline level without requiring a particular target expression level.

The methods disclosed herein result in a localized increase in HSPexpression, i.e., at or near a target site. In certain embodiments, themethods may further result in an increase in HSP expression levels atlocations remote to the target site. In certain of these embodiments,HSP expression levels may be elevated systemically, such that they canbe detected by a urine or blood draw from a subject.

In those embodiments of the methods disclosed herein that are directedto the treatment of a condition for which increased HSP expression isexpected to be beneficial, the condition may be selected from the groupconsisting of, for example, CVD (e.g., myocardial ischemia, acutemyocardial ischemia, chronic myocardial ischemia, MI,ischemia/reperfusion (I/R) injury, peripheral artery disease (PAD)),trauma, stroke, diabetes, atherosclerosis, asthma, other chronicinflammatory conditions, cancer, neurodegeneration, and variousneurological disorders. In certain embodiments, the condition may beassociated with decreased levels of HSP expression. In otherembodiments, the condition may not be associated with decreased levelsof HSP expression, but nonetheless may be expected to benefit fromincreased HSP expression.

In certain embodiments of the methods disclosed herein, the conditionbeing treated is CVD such as myocardial ischemia or MI. Myocardialischemia is one of the most common causes of heart failure, withsubsequent progression to MI. Even transient ischemia can lead tomyocardial necrosis and apoptosis. Myocardial reperfusion strategies aimto reduce morbidity and mortality by restoring tissue oxygenation, butmay in fact result in further cellular damage due to mitochondrialgeneration of reactive oxygen species. This phenomenon is known as I/Rinjury (Murphy Physiol Rev 88:581 (2008)). It is now recognized thatHSPs play an important role in protecting against I/R. In thoseembodiments where CVD is being treated, the target site may be locatedin or around the heart or a component of the circulatory system. Forexample, the target site may be located in or around a coronary artery.In these embodiments, thermal ablation may target the heart or a bloodvessel, or one or more nerves proximate to the heart or a blood vessel.In those embodiments that target CVD, thermal ablation may be carriedout in conjunction with percutaneous coronary intervention (PCI) orangioplasty.

In certain embodiments of the methods disclosed herein, the conditionbeing treated is PAD. For example, the methods may be used to treat lowextremity vascular arterial beds that have restricted blood flow due toatherosclerosis, including partial vessel occlusions that generateischemic conditions in the surround tissues. Induction of HSP expressionmay salvage tissue damage related to oxygen deprivation andintervention/reoxygenation-related cell death. In those embodimentswhere PAD is being treated, the target site may be located in or arounda peripheral blood vessel such as a peripheral artery, or in or around anerve proximate to a peripheral blood vessel. In those embodiments thattarget PAD, thermal ablation may be carried out in conjunction with PCI.

In certain embodiments of the methods disclosed herein, the conditionbeing treated is one where structural and functional preservation ofproteins may enhance cell survival. These conditions include, forexample, neurodegeneration, trauma, and stroke.

In certain embodiments of the methods disclosed herein, the conditionbeing treated is a condition associated with a transplanted graft,including for example a kidney graft. In these embodiments, the methodsmay be used to treat graft-versus-host disease. HSP70 expression hasbeen shown previously to improve tissue survival in animals undergoingischemia of transplanted kidneys (Perdrizet Transplant Proc 25:1670(1993)). In those embodiments where conditions associated withtransplanted grafts are being treated, the target site may be located inor around an artery feeding the transplanted graft such as a renalartery, or in or around a nerve proximate to the artery.

Treatment of a condition for which increased HSP expression is expectedto be beneficial may refer to preventing the condition, slowing theonset or rate of development of the condition, reducing the risk ofdeveloping the condition, preventing or delaying the development ofsymptoms associated with the condition, reducing or ending symptomsassociated with the condition, generating a complete or partialregression of the condition, or some combination thereof.

A target site may be located in any target tissue that can be subjectedto thermal ablation. As disclosed herein, thermal ablation of renalefferent nerves was shown to increase HSP expression at renal targetsites. Thermal ablation is likewise expected to result in a similarincrease in HSP expression levels in other tissue types. In certainembodiments of the methods disclosed herein, the target site may dependon the specific condition being treated. In certain embodiments, atarget site is located in a target tissue that is currently experiencingan HSP expression pattern or another characteristic associated with acondition for which increased HSP expression is expected to bebeneficial. In other embodiments, the target tissue has previouslyexperienced such expression patterns or other characteristics, or hasbeen deemed at risk for experiencing such expression patterns or othercharacteristics. In certain embodiments, a target site may exhibitdecreased levels of one or more HSPs. In certain embodiments, the targetsite may be located in a target tissue that is associated with thecondition being treated or symptoms thereof. For example, where thecondition being treated is cancer, the target site may be located at ornear a tumor site. Similarly, where the condition being treated isatherosclerosis, the target site may be located at or near a vesselexhibiting atherosclerotic plaque formation.

The methods disclosed herein may use any thermal ablation techniqueknown in the art, including the use of monopolar or bipolar RF energy,microwave energy, laser light or optical energy, ultrasound energy(e.g., intravascularly delivered ultrasound, extracorporeal ultrasound,HIFU), magnetic energy, direct heat energy, cryotherapeutic energy, or acombination thereof. Alternatively or in addition to these techniques,the methods may utilize one or more non-thermal neuromodulatorytechniques. For example, the methods may utilize sympathetic nervoussystem (SNS) denervation by removal of target nerves, injection oftarget nerves with a destructive drug or pharmaceutical compound, ortreatment of the target nerves with non-thermal energy modalities. Incertain embodiments, the magnitude or duration of the increase in HSPexpression may vary depending on the specific technique being used.

Thermal ablation may be carried out using any ablation device known inthe art. In certain embodiments, ablation may be carried out using adevice that is in direct or close contact with a target site. In certainof these embodiments, the ablation device may access the target site viaa minimally invasive route, for example via an intravascular pathwaysuch as a femoral, brachial, or radial point of entry. In theseembodiments, the ablation device and related components may have size,flexibility, torque-ability, kink resistance, or other characteristicssuitable for accessing narrow or difficult to reach portions of vessels.In other embodiments, the target site may be accessed using an invasivedirect access technique. In certain embodiments, the device may be usedexternally, i.e., without direct or close contact to the target site.

In certain embodiments, an ablation device for use in the methodsdisclosed herein may combine two or more energy modalities. The devicemay include both a hyperthermic source of ablative energy and ahypothermic source, making it capable of, for example, performing bothRF ablation and cryoablation. The distal end of the treatment device maybe straight (for example, a focal catheter), expandable (for example, anexpanding mesh or cryoballoon), or have any other configuration (e.g., ahelical or spiral coil). Additionally or alternatively, the treatmentdevice may be configured to carry out one or more non-thermalneuromodulatory techniques. For example, the device may comprise a meansfor diffusing a drug or pharmaceutical compound onto the targettreatment area (for example, a distal spray nozzle).

In those embodiments of the methods disclosed herein that include a stepof measuring HSP expression levels before or after thermal ablation,such measurement can be carried out using any method known in the art.For example, the measurement can utilize one or more capture ordetection agents that specifically bind HSP, such as an antibody orepitope-binding portion thereof, an HSP receptor or a portion thereof,or a nucleic acid complementary to all or a portion of the HSP mRNAsequence. FIGS. 1A-1B illustrates the use of a labeled antibody to bindand detect an HSP expressed on the artery wall (FIG. 1A) or a secretedHSP (FIG. 1B). In other examples, measurement of HSP expression mayutilize a detection agent that has a functional interaction with HSP.For example, the detection agent may be a substrate for an HSP or anenzyme or catalytic antibody for which HSP is a substrate. FIG. 1Cillustrates the use of a detection agent (represented by scissors) thatfunctions to cleave off a portion of an HSP. In still other examples,measurement of HSP may be carried out using imaging/spectroscopytechniques that allow HSP levels to be assessed in a non-invasive manneror by tissue sampling.

Capture or detection agents for use detecting and measuring HSPexpression may be in solution, or they may be immobilized on a surfacesuch as a bead, resin, or one or more surfaces of an ablation or othertreatment device, a component thereof, or a separate capture device.Examples of suitable resins include, for example, hydrophobic resins,cation/anion exchange resins (e.g., carboxymethyl,sulfopropyl/diethylamine), immobilized metal affinity chromatography(IMAC) resins, and polar chromatographic resins (e.g., silica gel). Inthose embodiments wherein capture or detection agents are immobilized onone or more surfaces of a treatment device, a component thereof, or aseparate capture device, the capture or detection agents may be on theoutside of the device, i.e., in direct contact with arterial blood orthe artery wall. In other embodiments, the capture or detection agentsmay be on an internal surface, such as the interior of a catheter or acapture compartment.

In certain embodiments, binding of HSP to a capture agent and/orinteraction of HSP with a detection agent results in a quantifiablesignal. This quantifiable signal may be, for example, a colorimetric,fluorescent, heat, energy, or electric signal. In certain embodiments,this signal may be transduced to an external visual output device (see,e.g., FIGS. 2A and 2B). In certain embodiments, a capture or detectionagent may be labeled, such as for example with an enzymatic orradioactive label. A capture or detection agent may be a bindingsubstrate for a secondary capture agent, such as a labeled antibody.

In certain embodiments, binding of HSP to a capture agent results in asignal that can be transduced to an external monitoring device. Forexample, binding of HSP to a capture or detection agent may be detectedusing a high sensitivity fluorescence technique such as a resonanceenergy transfer method (e.g., Forster resonance energy transfer,bioluminescence resonance energy transfer, or surface plasmon resonanceenergy transfer). FIG. 3 illustrates a quantum dot embodiment forgenerating a signal based on binding of HSP to an affinity ligandcapture agent (e.g., an antibody, peptide, small molecule drug, orinhibitor). Quantum dots are nanometer sized semiconductor crystals thatfluoresce when excited with the proper frequency of light (see, e.g.,Xing Nat Protoc 2:1152 (2007)). The emitted light is tuned by the sizeof the nanocrystal, and excitation frequencies range from near IR to UV.Dynamic visualization through skin has been demonstrated in animalsusing near IR radiation.

In certain embodiments of the methods disclosed herein, determination ofbaseline and/or post-ablation HSP expression or activity is carried outusing any immunoassay-based method. For example, HSP levels may bedetermined using an electrochemical immunosensor (see, e.g., FIGS. 13Aand 13B), which provides concentration-dependent signaling (see, e.g.,Centi Bioanalysis 1:1271 (2009); Rusling Analyst 135:2496 (2010)).Antibodies for use in an immunoassay-based determination of HSP level oractivity may be labeled or unlabeled.

In certain embodiments, determination of baseline and/or post-ablationHSP level or activity takes place in vivo. In these embodiments, thedetermination may be carried out using the same device that is used tocarry out ablation, or a component attached to the ablation device.Alternatively, determination of HSP level or activity may be carried outusing a separate device. In certain of these embodiments, the separatedevice is delivered to the ablation site via the same catheter used todeliver the treatment device. In other embodiments, determination ofbaseline and/or post-ablation HSP level or activity takes place ex vivo.

In certain embodiments, the interaction between HSP and a capture ordetection agent takes place at or near the ablation site. In certain ofthese embodiments, HSP binds to a capture or detection agent in thebloodstream or at the surface of the arterial wall. In theseembodiments, the capture or detection agent may be in solution (i.e., inthe bloodstream) or immobilized to a surface that is contact with thebloodstream and/or arterial wall. For example, a device or componentthereof in which a capture or detection agent is integrated may be aballoon coated with one or more detection molecules that inflates totouch the ablated artery wall (see, e.g., FIG. 4B). Captured biomarkersmay be detected in vivo, or the balloon-based device may be removed forbiomarker detection ex vivo.

In other embodiments, the interaction between HSP and a capture ordetection agent takes place away from the ablation site. For example,HSP may be removed from an ablation site and sequestered in a capturecompartment (see, e.g., FIG. 4A). Removal from the ablation site mayutilize one or more filters. For example, a first filter at the distalend of a capture compartment may be selected such that it allows passageof HSP into the capture compartment while preventing passage of otherbiomolecules. A second filter at a proximal end of the capture componentmay be selected such it prevents passage of HSP out of the capturecompartment while allowing blood to flow out of the capture compartment.Through the use of one or more filters, HSP may be concentrated withinthe capture compartment. Alternatively or in addition to the use offilters, one or more additional steps may be taken to concentrate HSP inthe capture compartment prior to or simultaneous with contacting of HSPwith capture or target agents. For example, HSP may be concentratedusing beads. In certain embodiments that utilize a capture compartment,HSP is contacted with capture or detection agents in the capturecompartment while still in the body. Alternatively, the capturecompartment may be removed from the body prior to contacting HSP withcapture or detection agents.

In one embodiment of the methods disclosed herein, an RF ablationcatheter is used as the treatment device. The RF ablation catheter isadvanced directly to a target site such as the renal artery or aspecific location within the renal artery via an intravascular pathway.In this embodiment, the RF ablation catheter comprises an elongate shafthaving a distal end and a plurality of electrodes (as the treatmentelements) coupled thereto. An introducer sheath may be used tofacilitate advancement of the device to the target site. Further, imageguidance, such as computed tomography (CT) fluoroscopy, intravascularultrasound (IVUS), optical coherence tomography (OCT), or other suitableguidance modality, or combinations thereof, may be used to aid theclinician's manipulation. In certain embodiments, image guidancecomponents (for example, IVUS or OCT) may be incorporated into thetreatment device itself. Once the treatment device is adequatelypositioned in contact or close proximity to the target site, the deviceis activated and RF energy is applied from the one or more electrodes,effectively ablating the arterial tissue. Ablation continues for apredetermined amount of time, or until visualization of the treatmentarea shows sufficient ablation. The RF ablation catheter is then removedfrom the patient. In those embodiments wherein HSP levels are measuredsubsequent to ablation, a sample for HSP expression measurement may beobtained using the device or a component attached thereto prior toremoval. In other embodiments, an introducer sheath may be left in placefollowing catheter removal, and a sampling device may be advanced to thetarget site.

In one embodiment, the methods disclosed herein utilize aneuromodulation system as set forth in FIG. 14 . This neuromodulationsystem 100 (“system 100”) can include a treatment device 102 forcarrying out ablation, an energy source or console 104 (e.g., an RFenergy generator, a cryotherapy console, etc.), and a cable 106extending between the treatment device 102 and the console 104. Thetreatment device 102 can include a handle 108, a therapeutic element110, and an elongated shaft 112 extending between the handle 108 and thetherapeutic element 110. The shaft 112 can be configured to locate thetherapeutic element 110 intravascularly or intraluminally at a treatmentlocation, and the therapeutic element 110 can be configured to provideor support therapeutically-effective neuromodulation at the treatmentlocation. In certain embodiments, the shaft 112 and the therapeuticelement 110 can be 3, 4, 5, 6, or 7 French or another suitable size.Furthermore, the shaft 112 and the therapeutic element 110 can bepartially or fully radiopaque and/or can include radiopaque markerscorresponding to measurements, e.g., every 5 cm. In certain embodiments,the treatment device comprises an optional monitoring or samplingcomponent for evaluating changes in HSP expression.

Intravascular delivery can include percutaneously inserting a guide wire(not shown) within the vasculature and moving the shaft 112 and thetherapeutic element 110 along the guide wire until the therapeuticelement 110 reaches the treatment location. For example, the shaft 112and the therapeutic element 110 can include a guide-wire lumen (notshown) configured to receive the guide wire in an over-the-wire (OTW) orrapid-exchange configuration (RX). Other body lumens (e.g., ducts orinternal chambers) can be treated, for example, by non-percutaneouslypassing the shaft 112 and therapeutic element 110 through externallyaccessible passages of the body or other suitable methods. In someembodiments, a distal end of the therapeutic element 110 can terminatein an atraumatic rounded tip or cap (not shown). The treatment device102 can also be a steerable or non-steerable catheter device configuredfor use without a guide wire.

The therapeutic element 110 can have a single state or configuration, orit can be convertible between a plurality of states or configurations.For example, the therapeutic element 110 can be configured to bedelivered to the treatment location in a delivery state and to provideor support therapeutically-effective neuromodulation in a deployedstate. In these and other embodiments, the therapeutic element 110 canhave different sizes and/or shapes in the delivery and deployed states.For example, the therapeutic element 110 can have a low-profileconfiguration in the delivery state and an expanded configuration in thedeployed state. In another example, the therapeutic element 110 can beconfigured to deflect into contact with a vessel wall in a deliverystate. The therapeutic element 110 can be converted (e.g., placed ortransformed) between the delivery and deployed states via remoteactuation, e.g., using an actuator 114 of the handle 108. The actuator114 can include a knob, a pin, a lever, a button, a dial, or anothersuitable control component. In other embodiments, the therapeuticelement 110 can be transformed between the delivery and deployed statesusing other suitable mechanisms or techniques.

In some embodiments, the therapeutic element 110 can include anelongated member (not shown) that can be configured to curve (e.g.,arch) in the deployed state, e.g., in response to movement of theactuator 114. For example, the elongated member can be at leastpartially helical/spiral in the deployed state. In other embodiments,the therapeutic element 110 can include a balloon (not shown) that canbe configured to be at least partially inflated in the deployed state.An elongated member, for example, can be well suited for carrying one ormore electrodes or transducers and for delivering electrode-based ortransducer-based treatment. A balloon, for example, can be well suitedfor containing refrigerant (e.g., during or shortly after liquid-to-gasphase change) and for delivering cryotherapeutic treatment. In someembodiments, the therapeutic element 110 can be configured forintravascular, transvascular, intraluminal, and/or transluminal deliveryof chemicals. For example, the therapeutic element 110 can include oneor more openings (not shown), and chemicals (e.g., drugs or otheragents) can be deliverable through the openings. For transvascular andtransluminal delivery, the therapeutic element 110 can include one ormore needles (not shown) (e.g., retractable needles) and the openingscan be at end portions of the needles.

The console 104 is configured to control, monitor, supply, or otherwisesupport operation of the treatment device 102. In other embodiments, thetreatment device 102 can be self-contained and/or otherwise configuredfor operation without connection to the console 104. As shown in FIG. 14, the console 104 can include a primary housing 116 having a display118. The system 100 can include a control device 120 along the cable 106configured to initiate, terminate, and/or adjust operation of thetreatment device 102 directly and/or via the console 104. In otherembodiments, the system 100 can include another suitable controlmechanism. For example, the control device 120 can be incorporated intothe handle 108. The console 104 can be configured to execute anautomated control algorithm 122 and/or to receive control instructionsfrom an operator. Furthermore, the console 104 can be configured toprovide feedback to an operator before, during, and/or after a treatmentprocedure via the display 118 and/or an evaluation/feedback algorithm124. In some embodiments, the console 104 can include a processingdevice (not shown) having processing circuitry, e.g., a microprocessor.The processing device can be configured to execute stored instructionsrelating to the control algorithm 122 and/or the evaluation/feedbackalgorithm 124. Furthermore, the console 104 can be configured tocommunicate with the treatment device 102, e.g., via the cable 106. Forexample, the therapeutic element 110 of the treatment device 102 caninclude a sensor (not shown) (e.g., a recording electrode, a temperaturesensor, a pressure sensor, or a flow rate sensor) and a sensor lead (notshown) (e.g., an electrical lead or a pressure lead) configured to carrya signal from the sensor to the handle 108. The cable 106 can beconfigured to carry the signal from the handle 108 to the console 104.

The console 104 can have different configurations depending on thetreatment modality of the treatment device 102. For example, when thetreatment device 102 is configured for electrode-based ortransducer-based treatment, the console 104 can include an energygenerator (not shown) configured to generate RF energy, pulsed RFenergy, microwave energy, optical energy, focused ultrasound energy(e.g., HIFU), direct heat energy, or another suitable type of energy. Insome embodiments, the console 104 can include an RF generator operablycoupled to one or more electrodes (not shown) of the therapeutic element110. When the treatment device 102 is configured for cryotherapeutictreatment, the console 104 can include a refrigerant reservoir (notshown) and can be configured to supply the treatment device 102 withrefrigerant, e.g., pressurized refrigerant in liquid or substantiallyliquid phase. Similarly, when the treatment device 102 is configured forchemical-based treatment, the console 104 can include a chemicalreservoir (not shown) and can be configured to supply the treatmentdevice 102 with the chemical. In certain embodiments, the therapeuticassembly may further include one or more thermoelectric devices (such asa Peltier device) for the application of heat or cold therapy. Anyenergy modality can either be used alone or in combination with otherenergy modalities. In some embodiments, the treatment device 102 caninclude an adapter (not shown) (e.g., a luer lock) configured to beoperably coupled to a syringe (not shown). The adapter can be fluidlyconnected to a lumen (not shown) of the treatment device 102, and thesyringe can be used, for example, to manually deliver one or morechemicals to the treatment location, to withdraw material from thetreatment location, to inflate a balloon (not shown) of the therapeuticelement 110, to deflate a balloon of the therapeutic element 110, or foranother suitable purpose. In other embodiments, the console 104 can haveother suitable configurations.

Upon delivery to a target site, the therapeutic assembly may be furtherconfigured to be deployed into a treatment state or arrangement fordelivering energy at the treatment site. In some embodiments, thetherapeutic assembly may be placed or transformed into the deployedstate or arrangement via remote actuation, for example, via an actuator,such as a knob, pin, or lever carried by the handle. In otherembodiments, however, the therapeutic assembly may be transformedbetween the delivery and deployed states using other suitable mechanismsor techniques. The monitoring system can provide feedback of parameterssuch as nerve activity to verify that the treatment assembly providedtherapeutically effective treatment.

In the deployed state, the therapeutic assembly can be configured tocontact an inner wall of a vessel and to carry out ablation or a seriesof ablations without the need for repositioning. For example, thetherapeutic element can be configured to create a single lesion or aseries of lesions, e.g., overlapping or non-overlapping. In someembodiments, the lesion or pattern of lesions can extend aroundgenerally the entire circumference of the vessel, but can still benon-circumferential at longitudinal segments or zones along a lengthwiseportion of the vessel. This can facilitate precise and efficienttreatment with a low possibility of vessel stenosis. In otherembodiments, the therapeutic element can be configured cause apartially-circumferential lesion or a fully-circumferential lesion at asingle longitudinal segment or zone of the vessel.

The proximal end of the therapeutic assembly is carried by or affixed tothe distal portion of the elongate shaft. A distal end of thetherapeutic assembly may terminate with, for example, a rounded tip orcap. Alternatively, the distal end of the therapeutic assembly may beconfigured to engage another element of the system or treatment device.For example, the distal end of the therapeutic assembly may define apassageway for engaging a guide wire (not shown) for the delivery of thetreatment device using over-the-wire (OTW) or rapid exchange (RX)techniques. If the treatment device and the sampling component are partof a single intervention device, the intervention device may include oneor more filters or testing media for in-system analysis of sample cells.

The monitoring system may include one or more energy sources (forexample, a coolant reservoir and/or an RF generator), which may behoused in a console or used as a stand-alone energy source. The consoleor energy source is configured to generate a selected form and magnitudeof energy for delivery to the target treatment site via the therapeuticassembly. A control mechanism such as a foot pedal may be connected tothe console to allow the operator to initiate, terminate, and optionallyto adjust various operational characteristics of the energy generator,including, but not limited to, power delivery. The system may alsoinclude a remote control device (not shown) that can be positioned in asterile field and operably coupled to the therapeutic assembly. In otherembodiments, the remote control device is configured to allow forselective activation of the therapeutic assembly. In other embodiments,the remote control device may be built into the handle assembly. Theenergy source can be configured to deliver the treatment energy via anautomated control algorithm and/or under the control of the clinician.In addition, the energy source or console may include one or moreevaluation or feedback algorithms to provide feedback to the clinicianbefore, during, and/or after therapy. The feedback can be based onoutput from the monitoring system.

The energy source can further include a device or monitor that mayinclude processing circuitry, such as one or more microprocessors, and adisplay. The processing circuitry can be configured to execute storedinstructions relating to the control algorithm. The energy source may beconfigured to communicate with the treatment device to control thetreatment assembly and/or to send signals to or receive signals from themonitoring system. The display may be configured to provide indicationsof power levels or sensor data, such as audio, visual, or otherindications, or may be configured to communicate the information toanother device. For example, the console may also be operably coupled toa catheter lab screen or system for displaying treatment information,including for example nerve activity before and after treatment, effectsof ablation or temperature therapy, lesion location, lesion size, etc.

The distal end of one embodiment of a treatment device includes a distalelectrode which is used to apply energy to the target cells (forexample, RF energy or ultrasound energy), although a plurality ofelectrodes can instead be used. Alternatively or additionally, thedistal end can include a light emitter such as a laser or LED diode, ora microwave antenna (not shown). The distal end can further include aradiopaque marker band for positioning the device.

The distal end of another embodiment of a treatment device includes acryoballoon, which is used to cool (remove energy from) the targetcells. In this embodiment, the distal end of the treatment deviceincludes an inner balloon and an outer balloon, which may help preventcoolant leaks from the cryoballoon. If additional energy modalities areused, the cryoballoon may include one or more electrodes, antennas, orlaser diodes, or LED diodes (not shown). The distal end may furtherinclude a radiopaque marker band for positioning the device.

One of ordinary skill in the art will recognize that the variousembodiments described herein can be combined. The following examples areprovided to better illustrate the disclosed technology and are not to beinterpreted as limiting the scope of the technology. To the extent thatspecific materials are mentioned, it is merely for purposes ofillustration and is not intended to limit the technology. One skilled inthe art may develop equivalent means or reactants without the exerciseof inventive capacity and without departing from the scope of thetechnology. It will be understood that many variations can be made inthe procedures herein described while still remaining within the boundsof the present technology. It is the intention of the inventors thatsuch variations are included within the scope of the technology.

EXAMPLES Example 1: Effect of RF Ablation and SNS Denervation on HSPLevels

Animals were broken into three groups of three animals each: naïve (notreatment), sham (catheterized but not ablated), and treated (subject toablation at 65° C. and 90 seconds using a spiral ablation catheterdevice). Left and right renal arteries and surrounding tissue sampleswere obtained by sampling tissue in the area of ablation at 10 minutes(“day 0”), 7 days, or 24 hours post-treatment. Slices from the center ofablation sites were removed for histopathological analysis, and theablation sites were cleaned up by removing any non-ablated tissue andpooled. Tissue was maintained during the dissection process usingRNALater® solution.

Pooled tissue samples were weighed and mixed under frozen conditions,and then added to round-bottomed tubes containing 2× stainless steelbeads (5 mm diameter) at room temperature. 900 μL QIAzol lysis reagentwas added to each tube, and the tissue was macerated using theTissueLyser II Adaptor Set with disruption at 30 Hz (3×2 minutes) torelease RNA. An additional 300 μL of lysis buffer was added to eachtube, and the disruption cycle was repeated (1×2 minutes at 30 Hz).Lysates were transferred to new Eppendorf tubes for mRNA isolation.

120 μl gDNA Eliminator Solution was added to each lysate sample, andtubes were shaken vigorously for 15 seconds. 180 μL of chloroform wasadded, and tubes were again shaken vigorously for 15 seconds. After 2-3minutes at room temperature, tubes containing homogenate werecentrifuged at 12,000×g for 15 minutes at 4° C. The centrifuge waswarmed to room temperature, and the upper aqueous phase was transferredto a new Eppendorf tube. An equal volume of 70% ethanol was added toeach tube with thorough mixing, and 700 μL of each sample wastransferred to an RNeasy Mini spin column in a 2 mL collection tube.Samples were centrifuged for 15 seconds at >8000×g (>10,000 rpm) at roomtemperature and flow-thru was discarded. The ethanol mixing and RNeasycentrifugation steps were repeated until all sample was used up. 700 μLof Buffer RWT was added to each spin column, followed by centrifugationfor 15 seconds at >8,000×g (>10,000 rpm) to wash the membrane. Flow-thruwas discarded, and 500 μL Buffer RPE was added each spin column,followed by centrifugation for 15 seconds at >8,000×g (>10,000 rpm).Flow thru was discarded, and 500 μl Buffer RPE was again added to eachspin column, followed by centrifugation for 2 minutes at >8,000×g(>10,000 rpm) to wash the membrane. RNeasy spin columns were placed in anew 2 mL collection tube and centrifuged at full speed for 1 minute. Thespin column was placed in a new 1.5 mL collection tube, 50 μL, RNasefree water was added directly to the spin column membrane, and RNAeluted was eluted by centrifugation for 1 minute at >8,000×g (>10,000rpm). This step was repeated using another 50 μL, of RNase free water.To ensure significance, A260 readings were verified to be greater than0.15. An absorbance of 1 unit at 260 nm corresponds to 44 μg of mRNA permL (A260=1=44 μg/mL) at neutral pH.

ABI High Capacity cDNA kits were used to convert mRNA to cDNA forquantitative real-time PCR (qPCR). PCR was performed in optical 384-wellplates, freshly prepared on the Eppendorf epMotion liquid handler. Finalreaction volume was 20 μL (4 μL, Taqman Assay+mixture of 6 μL, cDNA (3ng)+10 μL Universal Master Mix with UNG). Assays were performed toinclude +RT (reverse transcriptase) samples and, when appropriate, a −RTcontrol. Endogenous controls (×2) were run in triplicate and animalsamples were run only once for screening purposes. The real-time PCRprotocol included an initial step of 50° C. (2 minutes) to activate theDNA polymerase, denaturation by a hot start at 95° C. for 10 minutes,and 40 cycles of a two step program (denaturation at 95° C. for 15seconds for primer annealing/extension at 60° C. for 1 minute).Fluorescence data was collected at 60° C. Fluorescence was quantifiedwith the ABI PRISM 7900HT, and the resultant data was analyzed using SDSRQ Manager (1.2.1) Software (Sequence Detection System Software, AppliedBiosystems). Each biomarker was checked, and threshold and baseline wasadjusted to produce (in Δ Rn versus Cycle) an amplification curve of thetype suggested by Applied Biosystems in their “Relative QuantificationUsing Comparative Ct Getting Started Guide.” A calibrator was selectedfor calculation of the RQ (relative quantification). The calibrator wasbased on an average of 6× figures from the three naïve animals, left &right arteries, resulting in a numerical result of 1 for the naïve RQ.For calculation of the RQ for the standard deviation (SD) of the naïves,any other experimental animal was used as a calibrator (generally thefirst animal for Day 0 treated). RQ averages of animals (×3) in the sametreatment group were calculated for each point and for each biomarkerindividually, and plotted in bar graphs.

Renal NE and dopamine (DBN) levels in naïve, sham, and test animals wereevaluated at 10 minutes, 24 hours, and 7 days. Average kidney NEproduction post-ablation is shown in FIG. 5 .

The initial screen was carried out using 70 candidate biomarkers. HSPsexhibiting a significant increase in expression at 10 minutespost-ablation included DNAJA4 (FIG. 7 ), DNAJB1 (FIG. 8 ), HSPB1, CLU(FIG. 9A), HSPD1 (FIG. 10 ), and HSPH1 (FIG. 11A). HSPs exhibiting asignificant increase in expression at 24 hours post-ablation includedHSPD1 (FIG. 10 ), HSPH1 (FIG. 11B), HMOX1 (FIG. 12A), and HSPA5 (FIG. 6). HSPs exhibiting a significant increase in expression at 7 dayspost-ablation included HSPD1 (FIG. 10 ), HMOX1 (FIG. 12B), and CLU (FIG.9B).

HSP expression in response to thermal ablation will be further evaluatedin human vascular and neuronal cells (e.g., primary vascular cells suchas Human Coronary Artery Endothelial Cells (HCAEC) or Human CoronaryArtery Smooth Muscle Cells (HCASMC), neuronal cell lines). Expressioncan be evaluated by actually exposing cells to thermal ablation, or bysimply exposing the cells to one or more conditions that mimic in vivoablation (e.g., exposure to heat).

For experiments in which cells are exposed to heat, the cells aresubjected to elevated temperatures (e.g., 65-95° C.) for various timeintervals (e.g., 30-120 seconds) and allowed to recover at 37° C. HSPlevels and/or activity are measured at the end of the testing phase andat various timepoints during the recovery phase (e.g., every 1-5minutes).

Cell culture samples from various timepoints may be collected for HSPsecretomics analysis to evaluate release of HSP into culture.Secretomics experiments may be performed using iTRAQ methodology(Wiśniewski Arch Pathol Lab Med 132:1566 (2008)).

Treated and control cells from various timepoints are lysed and the cellcontents are analyzed for changes in HSP level or activity. Changes inlevel or activity of one or more control proteins may also be measured.Analysis may be carried out using the iTRAQ methodology. For example,samples may be diluted (depending on their initial concentration),digested with trypsin, and iTRAQ labeled using 8-Plex reagent. Theresultant complex protein digests are pooled together for MudPITanalysis. Each fraction is analyzed by LC-MS/MS, for example using anABSciex 5500 QTRAP® mass spectrometer, for acquisition of massspectroscopy data with the inclusion of iTRAQ quantitation data. Proteincharacterization for MS/MS and iTRAQ data can be performed using ABSciexProteinPilot software v 4.0 (Paragon algorithm).

Additional Examples

1. A method for increasing HSP expression level at or near a target sitein a subject, the method comprising performing a thermal ablationprocedure at or near the target site, wherein the thermal ablationprocedure results in an increase in the expression level of one or moreHSPs at or near the target site.

2. A method for treating a condition for which increased HSP expressionis expected to be beneficial in a subject, the method comprisingperforming a thermal ablation procedure, wherein the thermal ablationprocedure results in an increase in HSP expression level, and whereinthe increase in HSP expression level results in treatment of thecondition.

3. The method of either of examples 1 or 2 wherein HSP expression levelis increased within 30 minutes or less following the thermal ablationprocedure.

4. The method of either of examples 1 or 2 wherein HSP expression levelis increased within 10 minutes or less following the thermal ablationprocedure.

5. The method of either of examples 1 or 2 wherein HSP expression levelis increased at one or more timepoints selected from 10 minutes, 24hours, and 7 days following the ablation procedure.

6. The method of either of examples 1 or 2 wherein the thermal ablationprocedure is carried out using a modality selected from the groupconsisting of monopolar or bipolar radio frequency (RF) energy,microwave energy, laser or optical light energy, ultrasound energy,magnetic energy, direct heat energy, and cryotherapeutic energy.

7. The method of either of examples 1 or 2 wherein the target site is ina renal blood vessel of the human subject.

8. The method of example 7 wherein the thermal ablation proceduretargets renal sympathetic nerves.

9. The method of example 2, wherein the condition is selected from thegroup consisting of CVD, PAD, trauma, stroke, diabetes, atherosclerosis,asthma, other chronic inflammatory conditions, cancer,neurodegeneration, and various neurological disorders

10. The method of either of examples 1 or 2 wherein the thermal ablationprocedure results in an increase in expression of an HSP selected fromthe group consisting of HSPA5, DNAJA4, DNAJB1, CLU, HSPD1, HSPH1, HMOX1,and HSPB1.

11. A device for carrying out the method of any of examples 1-10.

Conclusion

The above detailed descriptions of embodiments of the technology are notintended to be exhaustive or to limit the technology to the precise formdisclosed above. Although specific embodiments of, and examples for, thetechnology are described above for illustrative purposes, variousequivalent modifications are possible within the scope of thetechnology, as those skilled in the relevant art will recognize. Forexample, while steps are presented in a given order, alternativeembodiments may perform steps in a different order. The variousembodiments described herein may also be combined to provide furtherembodiments. All references cited herein are incorporated by referenceas if fully set forth herein.

From the foregoing, it will be appreciated that specific embodiments ofthe technology have been described herein for purposes of illustration,but well-known structures and functions have not been shown or describedin detail to avoid unnecessarily obscuring the description of theembodiments of the technology. Where the context permits, singular orplural terms may also include the plural or singular term, respectively.

Moreover, unless the word “or” is expressly limited to mean only asingle item exclusive from the other items in reference to a list of twoor more items, then the use of “or” in such a list is to be interpretedas including (a) any single item in the list, (b) all of the items inthe list, or (c) any combination of the items in the list. Additionally,the term “comprising” is used throughout to mean including at least therecited feature(s) such that any greater number of the same featureand/or additional types of other features are not precluded. It willalso be appreciated that specific embodiments have been described hereinfor purposes of illustration, but that various modifications may be madewithout deviating from the technology. Further, while advantagesassociated with certain embodiments of the technology have beendescribed in the context of those embodiments, other embodiments mayalso exhibit such advantages, and not all embodiments need necessarilyexhibit such advantages to fall within the scope of the technology.Accordingly, the disclosure and associated technology can encompassother embodiments not expressly shown or described herein.

We claim:
 1. A method of treating a human patient diagnosed with acancerous tumor, the method comprising: intravascularly positioning acatheter carrying a neuromodulation treatment device adjacent to atarget sympathetic nerve innervating tissue proximate the tumor in thepatient; and delivering energy to the target sympathetic nerve via theneuromodulation assembly to reduce sympathetic nerve activity in thetarget sympathetic nerve, wherein reducing sympathetic nerve activity inthe target sympathetic nerve results in a therapeutically beneficialtreatment of the tumor.
 2. The method of claim 1, wherein reducingsympathetic nerve activity in the target sympathetic nerve results in anincrease in an HSP expression level, and wherein after delivering energyto the target sympathetic nerve, the method further comprises:quantifying the HSP expression level; and comparing the HSP expressionlevel to a pre-determined threshold expression level for the HSP,wherein the HSP expression level is beneficial in the treatment of thetumor if the HSP expression level is greater than the pre-determinedthreshold expression level.
 3. The method of claim 2, wherein the HSPexpression level includes one or more of an HSP mRNA level, an HSPprotein level, a level of secreted HSP protein and a level of HSPactivity.
 4. The method of claim 1, wherein the cancerous tumor isidentified as a cancer for which increased HSP expression is expected tobe beneficial.
 5. The method of claim 1, wherein the cancerous tumor isin the kidney of the patient, and wherein the neuromodulation treatmentdevice is positioned within the renal artery.
 6. The method of claim 1,wherein reducing sympathetic nerve activity in the target sympatheticnerve comprises at least partially ablating the target sympatheticnerve.
 7. The method of claim 1, wherein reducing sympathetic nerveactivity in the target sympathetic nerve comprises increasing an HSPexpression level in the tissue proximate the tumor in the patient. 8.The method of claim 1, wherein a therapeutically beneficial treatment ofthe tumor includes one or more of slowing the onset or rate ofdevelopment of the tumor, reducing the risk of spread of the tumor,preventing or delaying the development of symptoms associated with thetumor, reducing or ending symptoms associated with the tumor, generatinga complete or partial regression of the tumor, or a combination thereof.